Cryopreservation of Plagiognathops microlepis sperm |
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| Institution: | 1. State Key Laboratory of Biocontrol, Institute of Aquatic Economic Animals and the Guangdong Province Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China;2. Nanhai Bairong Improved Aquatic Seed CO.,TLD, Foshan, China;3. Guangdong Provincial Fishery Germplasm Conservation Center, Guangzhou, China;1. Faculty of Fisheries and Protection of Waters, University of South Bohemia in Ceske Budejovice, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zatisi 728/II, Vodnany 389 25, Czech Republic;2. Scientific-Research Institute of Biology, V.N. Karazin Kharkiv National University, Svobody Sq. 4, Kharkiv 61022, Ukraine;1. Fisheries Department, Faculty of Natural Resources, Urmia University, Urmia, Iran;2. Young Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran;3. Rajaei Sturgeon Hatchery Center, Sari, Iran;4. Fisheries Department, Faculty of Agriculture and Natural Resources, Persian Gulf University, Bushehr, Iran;5. Medicine Laboratory, Alavi Educational and Treatment Center, Ardabil University of Medical Sciences, Ardabil, Iran;6. Biotechnology Laboratory, Faculty of Agriculture and Natural Resources, Tehran University, Karaj, Iran;1. School of Marine Sciences, Ningbo University, Ningbo, Zhejiang 315211, People''s Republic of China;2. The Sperm Laboratory, College of life Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, People''s Republic of China;1. The Key Laboratory of Applied Marine Biotechnology by the Ministry of Education, College of Ocean, Ningbo University, Ningbo 315211, China;2. The Sperm Laboratory, College of Life Sciences, Zhejiang University, 866 Yu Hang Tang Road, Hangzhou, Zhejiang 310058, China;3. Zhejiang Institute of Freshwater Fisheries, Huzhou, Zhejiang 313001, China |
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| Abstract: | Sperm was collected from cultured male fish and cryopreserved in 0.25 ml straws for the study of sperm cryopreservation. Different parameters were evaluated, including extender, dilution ratio, cryoprotectant type and concentration, equilibrium time, cooling height (in a two-step cooling protocol), and thawing temperature. The optimum result was obtained when the sperm was diluted at a 1:7 ratio in D-16 with 5% DMSO as a cryoprotectant, equilibrated for 20 min, held at 3 cm above liquid nitrogen for 10 min, and then stored in liquid nitrogen. After thawing in a water bath at 40 °C, the percentage of motile cells and fertilization rates of frozen-thawed sperm were 35.33 ± 2.52% and 39.00 ± 4.58%, respectively, while the corresponding rates for fresh sperm were 87.67 ± 3.06% and 88.67 ± 4.62%. We also used a programmed cooling protocol in which temperature was decreased from 4 °C to −80 °C by a rate of 30 °C/min, and then straws (0.25 ml) were placed above the surface of liquid nitrogen for 2 min before being stored in liquid nitrogen. This protocol provided a post-thaw activation rate of 36.67 ± 4.77%. Further parametric optimization is required to improve the quality of frozen-thawed sperm. |
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| Keywords: | Sperm cryopreservation Cryoprotectant |
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